Aykut, Ayse Ozlem; Atilgan, Ali Rana; Atilgan, Canan
Designing Molecular Dynamics Simulations to Shift Populations of the Conformational States of Calmodulin
PLOS COMPUTATIONAL BIOLOGY, 9 Art. No. e1003366, DEC 2013

We elucidate the mechanisms that lead to population shifts in the conformational states of calcium-loaded calmodulin (Ca2+-CaM). We design extensive molecular dynamics simulations to classify the effects that are responsible for adopting occupied conformations available in the ensemble of NMR structures. Electrostatic interactions amongst the different regions of the protein and with its vicinal water are herein mediated by lowering the ionic strength or the pH. Amino acid E31, which is one of the few charged residues whose ionization state is highly sensitive to pH differences in the physiological range, proves to be distinctive in its control of population shifts. E31A mutation at low ionic strength results in a distinct change from an extended to a compact Ca2+-CaM conformation within tens of nanoseconds, that otherwise occur on the time scales of microseconds. The kinked linker found in this particular compact form is observed in many of the target-bound forms of Ca2+-CaM, increasing the binding affinity. This mutation is unique in controlling C-lobe dynamics by affecting the fluctuations between the EF-hand motif helices. We also monitor the effect of the ionic strength on the conformational multiplicity of Ca2+-CaM. By lowering the ionic strength, the tendency of nonspecific anions in water to accumulate near the protein surface increases, especially in the vicinity of the linker. The change in the distribution of ions in the vicinal layer of water allows N- and C- lobes to span a wide variety of relative orientations that are otherwise not observed at physiological ionic strength. E31 protonation restores the conformations associated with physiological environmental conditions even at low ionic strength. Author Summary Calmodulin (CaM) is involved in calcium signaling pathways in eukaryotic cells as an intracellular Ca2+ receptor. Exploiting pH differences in the cell, CaM performs a variety of functions by conveniently adopting different conformational states. We aim to reveal pH and ionic strength (IS) dependent shifts in the populations of conformational substates by modulating electrostatic interactions amongst the different regions of the protein and with its vicinal water. For this purpose, we design extensive molecular dynamics simulations to classify the effects that are responsible for adopting different conformations exhibited in the ensemble of NMR structures reported. Lowering the IS or pH, CaM experiences higher inter-lobe orientational flexibility caused by extreme change in the non-specific ion distribution in the vicinal solvent. Amongst the titratable groups sensitive to pH variations, E31 is unique in that its protonation has the same effect on the vicinal layer as increasing the IS. Furthermore, E31A mutation causes a large, reversible conformational change compatible with NMR ensemble structures populating the linker-kinked conformations. The mutation in the N lobe, at a significant distance, both modulates the electrostatic interactions in the central linker and alters the EF-hand helix orientations in the C lobe.

DOI:10.1371/journal.pcbi.1003366

Find full text with Google Scholar.